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Worthington 骨骼组织分离指南

发布者:艾美捷科技    发布时间:2022-05-31     
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原代细胞分离中,针对骨骼组织涉及到软骨细胞、成骨细胞、骨和骨髓基质细胞、股骨近端等等的解离通常会使用酶,如何选择合适的酶来完成对应骨骼组织的解离,以及原代细胞采集的常规操作, 艾美捷针对骨骼解离分种属、不同的细胞、酶组合推荐,推荐浓度、培养基以及参考文献,详细汇总指南:tissue-worthington.png 


本指南总结了Worthington本技术指南描述了标准实验室程序; 为建立细胞分离方案提供了一种合乎逻辑的实验方法; 并列出了许多组织特异性参考资料,以帮助开发有效的方法。更多其他组织的解离指南,点击查看:各组织分离指南

 

骨骼
种属详情细胞名称酶组合推荐,含推荐浓度培养基参考文献
Bovine软骨细胞 / ChondrocytesCollagenase Type 2: 0.4%DMEMBuschmann, M., Gluzband, Y., Grodzinsky, A., and Hunziker, E.: Mechanical Compression Modulates Matrix Biosynthesis in Chondrocyte/Agarose Culture, J Cell Sci 108, 1497, 1995
Chick, day old成骨细胞 / OsteoblastsTrypsin: 0.03%DMEMGay, C., Lloyd, Q., and Gilman, V.: Characteristics and Culture of Osteoblasts Derived From Avian Long Bone, In Vitro Cell Dev Biol 30A, 379, 1994
Chick, Peterson/Arbor Acre, male, 4 weeks old (Gallus domesticus)软骨细胞 / ChondrocytesTrypsin: 0.25%Ham's F12Rosselot, G., Reginato, A.M., and Leach, R.M.: Development of a Serum-Free System to Study the Effect of Growth Hormone and Insulinlike Growth Factor-1 on Cultured Postembryonic Growth Plate Chondrocytes, In Vitro Cell Dev Biol 28A, 235, 1992
Chick, embryo椎骨成软骨细胞 / Vertebrae chondroblastsTrypsin: 0.25%Simm's, CMFSchiltz, J.R., Mayne R., and Holtzer, H.: The Synthesis of Collagen and Glycosaminoglycans by Dedifferentiated Chondroblasts in Culture, Differentiation 1, 97, 1973
Human骨-软骨-基质细胞 / Bone-cartilage-stromalCollagenase Type 2: 0.75%HBSSAgarwal, S., Loder, S., Sorkin, M., Li, S., Shrestha, S., Zhao, B., Mishina, Y., James, A. and Levi, B.: Analysis of Bone-Cartilage-Stromal Progenitor Populations in Trauma Induced and Genetic Models of Heterotopic Ossification., Stem Cells 34, 1692-701, 2016
Human成骨细胞 / OsteoblastsCollagenase Type 2: 0.2%;
Trypsin: 0.25%-1.0%
DMEMTaylor, S., Shah, M. and Orriss, I.: Generation of Rodent and Human Osteoblasts., Bonekey Rep 3, 585, 2014
Human成骨细胞 / OsteoblastsCollagenase Type 3: 0.2%MEMKode, A.,Manavalan1, J.,Mosialou1, I., Bhagat G., Rathinam, C, Luo, N et al.: Leukemogenesis Induced by an Activating β-catenin Mutation in Osteoblasts, Nature 506, 240, 2014
Human, 22-73 year成骨细胞 / OsteoblastsTrypsin: 0.5%Basal MediumKneser U, Voogd A, Ohnolz J, Buettner O, Stangenberg L, Zhang YH, Stark GB, Schaefer DJ: Fibrin gel-immobilized primary osteoblasts in calcium phosphate bone cement: in vivo evaluation with regard to application as injectable biological bone substitute, Cells Tissues Organs 179, 158-69, 2005
Human, 60+ year骨细胞,成骨细胞 / Bone Cells, OsteoblastsCollagenase Type 2: 200-250 u/mlDMEMChen X., Qian H., Neff L., Satomura K., and Horowitz M.: Thy-1 antigen Expression by Cells in the Osteoblast Lineage, J Bone Miner Res 14, 362, 1999
Human成骨细胞 / OsteoblastsTrypsin: 0.1%DMEMMeikle, M., Boyd, S., Hembry, R., Compston, J., Croucher, P., and Reynolds, J.: Human Osteoblasts in Culture Synthesize Collagenase and Other Matrix Metalloproteinases in Response to Osteotropic Hormones and Cytokines, J Cell Sci 103, 1093, 1992
Human成骨细胞 / OsteoblastsCollagenase Type 4: 250 u/mlDMEMFedarko, N.S., Termine, J.D., Young, M.F. and Robey, P.G.: Temporal Regulation of Hyaluronan and Proteoglycan Metabolism by Human Bone Cells in Vitro, J Biol Chem 265, 12200, 1990
Mouse骨和骨髓基质细胞 / Bone and bone marrow stromalStemxyme 1: 0.1%;
Neutral Protease: 0.1%
PBSMohrin, D., Koukos, G., Alabdulaaly, L., Paw, J., Hake, K., Kolumam, G., Bouxsein, M., Baron, R., Kutskova, Y., Freund, A., Liu, J., Zavala-Solorio, J., Bhargava, S., Maxwell, Trumble, J., Brito, A., Hu, D. and Brooks, D.: Inhibition of Longevity Regulator PAPP-A Modulates Tissue Homeostasis via Restraint of Mesenchymal Stromal Cells.,Aging Cell Vol. 20,, , e13313, 2021
Mouse骨和骨髓基质细胞 / Bone and bone marrow stromaStemxyme 1: 0.1%;
Neutral Protease: 0.1%
Media 199Baryawno, N., Przybylski, D., Kowalczyk, M., Kfoury, Y., Severe, N., Gustafsson, K., Kokkaliaris, K., Mercier, F., Tabaka, M., Hofree, M., Dionne, D., Papazian, A., Lee, D., Ashenberg, O. and Subramanian, A.: A Cellular Taxonomy of the Bone Marrow Stroma in Homeostasis and Leukemia., Cell 177, 1915-1932.e16, 2019
Mouse成骨细胞 / OsteoblastCollagenase Type 1: 0.3%HBSSWang, W., Majihail, G., Lui, C. and Zhou, L.: Osteoblast Sorting and Intracellular Staining of CXCL12., Bio Protoc 8, , 2018
Mouse骨-软骨-基质细胞 / Bone-cartilage-stromalCollagenase Type 2: 0.75%HBSSAgarwal, S., Loder, S., Sorkin, M., Li, S., Shrestha, S., Zhao, B., Mishina, Y., James, A. and Levi, B.: Analysis of Bone-Cartilage-Stromal Progenitor Populations in Trauma Induced and Genetic Models of Heterotopic Ossification., Stem Cells 34, 1692-701, 2016
Mouse骨基质细胞 / Bone stromalCollagenase Type 1: 0.3%DMEMIgarashi, H., Akahoshi, N., Ohto-Nakanishi, T., Yasuda, D. and Ishii, S.: The Lysophosphatidic Acid Receptor LPA4 Regulates Hematopoiesis-Supporting Activity of Bone Marrow Stromal Cells., Sci Rep 5, 11410, 2015
Mouse成骨细胞 / OsteoblastsCollagenase Type 1: 1.0%;
Neutral Protease: 1.6%
MEMLee, D., Choung, H., Kim, H., Gronostajski, R., Yang, Y., Ryoo, H., Lee, Z., Kim, H., Cho, E. and Park, J.: NFI-C Regulates Osteoblast Differentiation Via Control of Osterix Expression., Stem Cells 32, 2467-79, 2014
Mouse, 2-5 day成骨细胞 / OsteoblastsCollagenase Type 2: 0.2%;
Trypsin: 0.25%-1.0%
DMEMTaylor, S., Shah, M. and Orriss, I.: Generation of Rodent and Human Osteoblasts., Bonekey Rep 3, 585, 2014
Mouse成骨细胞 / OsteoblastsCollagenase Type 3: 0.2%MEMKode, A.,Manavalan1, J.,Mosialou1, I., Bhagat G., Rathinam, C, Luo, N et al.: Leukemogenesis Induced by an Activating β-catenin Mutation in Osteoblasts, Nature 506, 240, 2014
Mouse, 4-22 month骨细胞 / OsteocytesCollagenase Type 1: 300 u/mlMEMStern, A., Stern, M., Van Dyke, M., Jahn, K., Prideaux, M. and Bonewald, L.: Isolation and Culture of Primary Osteocytes from the Long Bones of Skeletally Mature and Aged Mice., Biotechniques 52, 361, 2012
Mouse骨内膜细胞 / Endosteal cellsCollagenase Type 1: 0.3%DMEMNakamura, Y., Arai, F., Iwasaki, H., Hosokawa, K., Kobayashi, I., Gomei, Y., Matsumoto, Y., Yoshihara, H. and Suda, T.: Isolation and Characterization of Endosteal Niche Cell Populations that Regulate Hematopoietic Stem Cells., Blood 116, 1422, 2010
Mouse骨髓 / Bone marrowCollagenase: 0.2%DMEMMorikawa, S., Mabuchi, Y., Kubota, Y., Nagai, Y., Niibe, K., Hiratsu, E., Suzuki, S., Miyauchi-Hara, C., Nagoshi, N., Sunabori, T., Shimmura, S., Miyawaki, A., Nakagawa, T., Suda, T., Okano, H. and Matsuzaki, Y.: Prospective Identification, Isolation, and Systemic Transplantation of Multipotent Mesenchymal Stem Cells in Murine Bone Marrow., J Exp Med 206, 2483-96, 2009
Mouse, male, 6-8 week old破骨细胞 / OsteoclastsCollagenase Type 3: 0.1%DMEMSakai, E., Miyamoto, H., Okamoto, K., Kato, Y., Yamamoto, K., and Sakai, H.: Characterization of Phagosomal Subpopulations Along Endocytic Routes in Osteoclasts and Macrophages, J. Biochem. 130, 823, 2001
Mouse, 3-5 day, 6-8 week骨细胞,成骨细胞 / Bone Cells, OsteoblastsCollagenase Type 2: 200-250 u/mlDMEMChen X., Qian H., Neff L., Satomura K., and Horowitz M.: Thy-1 antigen Expression by Cells in the Osteoblast Lineage, J Bone Miner Res 14, 362, 1999
Mouse, BALB/c成骨细胞样细胞,基质细胞系,造血母细胞 / Osteoblast-like Cells,Stromal Cell Lines,Hematopoietic Blast CellsTrypsin: 0.1%Eagle's MEMTakanashi, H., Matsuishi, T., and Yoshizato, K.: Establishment and Characterization of Stromal Cell Lines That Support Differentiation of Murine Hematopoietic Blast Cells into Osteoblast-like Cells, In Vitro Cell Dev Biol 30A, 384, 1994
Mouse, Swiss-Webster新生儿骨 / Neonatal boneCollagenase Type 2: 0.20%Tris-buffered salineChen,T. and Feldman,D.: Regulation of 1,25-Dihydroxyvitamin D3 Receptors in Cultured Mouse Bone Cells, J Biol Chem 256, 5561, 1981
Rat, 2 day成骨细胞 / OsteoblastsCollagenase Type 2: 0.1%;
Trypsin: 0.25%
DMEMJeon, J., Lee, M. and Yang, H.: Differentiated Osteoblasts Derived Decellularized Extracellular Matrix to Promote Osteogenic Differentiation., Biomater Res 22, 4, 2018
Rat, Wistar, 250g股骨近端 / Proximal femur stemCollagenase Type 1: 0.075%HBSSJacobs, F., Gijsen, H., Van de Vyver, M. and Ferris, W.: Vanadate Impedes Adipogenesis in Mesenchymal Stem Cells Derived from Different Depots within Bone., Front Endocrinol 7, 108, 2016
Rat, 2-3 day成骨细胞 / OsteoblastsCollagenase Type 2: 0.2%;
Trypsin: 0.25%-1.0%
DMEMTaylor, S., Shah, M. and Orriss, I.: Generation of Rodent and Human Osteoblasts., Bonekey Rep 3, 585, 2014
Rat, fetal, 21 days of gestation卡尔瓦利亚 / CalvariaCollagenase: 0.2%MEMOwen, T., Aronow, M., Shalhoub, V., Barone, L., Wilming, L., Tassinari, M., Kennedy, M., Pockwinse, S., Lian, J., and Stein, G.: Progressive Development of the Rat Oseoblast Phenotype In Vitro: Reciprocal Relationships in Expression of Genes Associated with Osteoblast Proliferation and Differentiation During Formation of the Bone Extracellular Matrix, J Cell Physiol 143, 420, 1990
Rat, newborn成骨细胞样细胞 / Osteoblastlike cellsCollagenase Type 2: 0.3%MEMErnst, M., and Froesch, E.: Osteoblastlike Cells in a Serum-Free Methylcellulose Medium Form Colonies: Effects of Insulin and Insulinlike Growth Factor I, Calcif Tissue Int 40, 27, 1987
Rat, fetus, 17-21 day卡尔瓦利亚 / CalvariaCollagenase: 0.01%-0.6%Tris-buffered salinePeck, W., Birge, S., and Fedak, S.: Bone Cells: Biochemical and Biological Studies After Enzymatic Isolation, Science 146, 1476, 1964


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